Cryopreservation and metabolic profiling analysis of Arabidopsis T87 suspension-cultured cells.
We established a simple cryopreservation protocol for Arabidopsis T87 cells using an encapsulation-dehydration method. T87 cells were encapsulated into alginate beads containing 2 M glycerol and 0.4 M sucrose. Alginate beads containing T87 cells were dehydrated with silica gel for 2 h (to c. 0.7 g water per g dry weight followed by immersed in LN. After rewarming at 35 degree C for 3 min and 1-d incubation under continuous illumination at 22 degree C, cryopreserved T87 cells exhibited considerable regrowth. Exponentially-grown 7-d-old T87 cells regrew more vigorously (86% of control) than 14-d-old cells after cryopreservation without preculture in medium containing 0.3 M sucrose. Genetic stability of cryopreserved T87 cells was demonstrated by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) and principal component analysis (PCA). Transformed T87 cells were cryopreserved using established protocols, and GUS expression was maintained within a 2-fold variance. These results indicate that cryopreservation of T87 cells is useful for comprehensive metabolomics research and for the large scale collection of transformed cultured cell lines for functional genomics research.
Ogawa
Yoichi
Y
Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.
Suzuki
Hideyuki
H
Sakurai
Nozomu
N
Aoki
Koh
K
Saito
Kazuki
K
Shibata
Daisuke
D
D016960:Agrobacterium tumefaciens; D017360:Arabidopsis / Q000254:growth & development / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D015925:Cryopreservation; D003890:Desiccation; D008401:Gas Chromatography-Mass Spectrometry; D005822:Genetic Vectors; D005966:Glucuronidase / Q000378:metabolism; D055442:Metabolome; D008863:Microspheres; D014170:Transformation, Genetic
D005966:Glucuronidase